Superior<i>In vivo</i>Efficacy of Afucosylated Trastuzumab in the Treatment of HER2-Amplified Breast Cancer

Junttila, Teemu T.; Parsons, Kathryn L.; Olsson, Christine; Lu, Yanmei; Xing, Yan; Thériault, Julie M.; Crocker, Lisa; Pabonan, Oliver; Baginski, Tomasz; Meng, Gloria; Tótpál, Klára; Kelley, Robert F.; Sliwkowski, Mark X.

doi:10.1158/0008-5472.can-09-3704

Cited by 199 publications

(179 citation statements)

References 37 publications

(44 reference statements)

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“…3. No effect of Ox or fucosylation degree of IgG was measured on FcγRI binding, confirming earlier results in the literature on FcγRI binding for oxidized IgG1 20 and fucosylation 17, 18.…”

Section: Resultssupporting

confidence: 89%

“…Furthermore, fucosylation levels of the antibody have an impact on the binding to FcγRIIIa and FcγRIIIb, whereas no differences in binding were measured on any of the other Fc receptors. These results are in agreement with the literature 13, 16, 17, 18, 19. None of the cited references studied the effect of DG on fraction bound at neutral pH, and we have demonstrated that there is a significant impact.…”

Section: Discussionsupporting

confidence: 93%

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Rapid screening of IgG quality attributes – effects on Fc receptor binding

Geuijen¹,

Oppers-Tiemissen

Egging

et al. 2017

FEBS Open Bio

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The interactions of therapeutic antibodies with fragment crystallizable γ (Fcγ) receptors and neonatal Fc receptors (FcRn) are measured in vitro as indicators of antibody functional performance. Antibodies are anchored to immune cells through the Fc tail, and these interactions are important for the efficacy and safety of therapeutic antibodies. High‐throughput binding studies on each of the human Fcγ receptor classes (FcγRI, FcγRIIa, FcγRIIb, FcγRIIIa, and FcγRIIIb) as well as FcRn have been developed and performed with human IgG after stress‐induced modifications to identify potential impact in vivo. Interestingly, we found that asparagine deamidation (D‐N) reduced the binding of IgG to the low‐affinity Fcγ receptors (FcγRIIa, FcγRIIb, FcγRIIIa, and FcγRIIIb), while FcγRI and FcRn binding was not impacted. Deglycosylation completely inhibited binding to all Fcγ receptors, but showed no impact on binding to FcRn. On the other hand, afucosylation only impacted binding to FcγRIIIa and FcγRIIIb. Methionine oxidation at levels below 7%, multiple freeze/thaw cycles and short‐term thermal/shake stress did not influence binding to any of the Fc receptors. The presence of high molecular weight species, or aggregates, disturbed measurements in these binding assays; up to 5% of aggregates in IgG samples changed the binding and kinetics to each of the Fc receptors. Overall, the screening assays described in this manuscript prove that rapid and multiplexed binding assays may be a valuable tool for lead optimization, process development, in‐process controls, and biosimilarity assessment of IgGs during development and manufacturing of therapeutic IgGs.

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Section: Resultssupporting

confidence: 89%

Section: Discussionsupporting

confidence: 93%

Rapid screening of IgG quality attributes – effects on Fc receptor binding

Geuijen¹,

Oppers-Tiemissen

Egging

et al. 2017

FEBS Open Bio

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“…It is known that mouse FcγRs and FcRn efficiently bind human IgG subclasses, but the affinity is lower than that of the human versions, so humanized antibody Fc cannot effectively activate NK cells and macrophages of BALB/c nude mice compared with mouse IgG subclasses. 23 However, the ADCC function of MBS301 can be observed in the humanized mice (hu-CD34 NSG mice) study (Figure 7c); there was a slight visible difference between the MBS301 and MBS301 + F groups in tumor inhibition and the recurrence ratio of MBS301 was lower than that of MBS301 + F. 24 Still, the content of humanized NK cells was much lower in hu-CD34 NSG mice than in human. 25 The NK cells could account for 5–20% in human peripheral blood mononuclear cells; thus, significant ADCC function for killing cancer cells could be observed for MBS301 when used as a treatment in humans.…”

Section: Discussionmentioning

confidence: 93%

Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

Huang

Liu

et al. 2018

mAbs

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MBS301, a glyco-engineered bispecific anti-human epidermal growth factor receptor 2 (HER2) antibody with a typical IgG1 monoclonal antibody structure, was developed through dual-cell expression and in vitro assembling process. MBS301 consists of two half antibodies engineered from trastuzumab and pertuzumab, respectively. Integrity and purity profiles of MB301 indicated that the heterodimerization of the two half antibodies was successful. The high and similar melting temperatures (Tm1,72.0°C and Tm2, 84.8°C) of MBS301 compared with those of its parental monoclonal antibodies trastuzumab and pertuzumab (in-house made T-mab and P-mab, respectively) revealed its structural compactness. With computer-modeling experiments and Biacore binding and competition kinetics studies, the binding stoichiometry between MBS301 and HER2-ECD was determined to be 1:1 and the two arms of MBS301 were shown to bind to domains II and IV of HER2-ECD antigen simultaneously. MBS301 displayed synergistic bioactivities as the combination of T-mab and P-mab in vitro in multiple cancer cell lines and in vivo in xenograft mouse model studies, and showed more effective activity than T-mab or P-mab used individually. Moreover, fucose-knockout dramatically increased MBS301’s binding affinity to low affinity FcγRIIIa allotype 158F (KD = 2.35 × 10−7M) to near the high affinity level of allotype V158 (KD = 1.17 × 10−7M). This resulted in far more effective ADCC activity of MBS301 than the combination of T-mab and P-mab in killing HER2-positive cancer cells. Hence, a novel fully afucosylated anti-HER2 bispecific antibody with improved antitumor activities was generated and shown to have the potential to be used for treating HER2-positive but trastuzumab-resistant solid tumors.

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“… 132 Humanized IgG1 Bemarituzumab /FPA144FGFR2bafucosylatedFUT8 −/− CHO cells (Potelligent® Technology)Enhanced tumour growth inhibition in FGFR2-overexpressing gastric cancer xenograft mouse model over isotype control(b)Humanized IgG1 Afucosylated TrastuzumabHER2afucosylatedFUT8 −/− CHO cells (Potelligent® Technology)Enhanced tumor growth inhibition over fucosylated trastuzumab in KPL-4 xenografts in human FcγRIIIα miceJunttila et al. 133 Humanized IgG1 TrasGEX/ GT-MAB7.3-GEX/Glycooptimized Trastuzumab-GEXHER2ReducedHuman glycoengineered production cell lines (GlycoExpress technology)Potent Her2 + BT474 tumor growth inhibition in nude mice but exhibited similar tumor volume and number of regression between fucosylated and reduced fucosylated antibodies(c)Humanized IgG1 Lumretuzumab /RG7116/RO5479599/ GE-HuMAb-HER3HER3ReducedCoexpression with GnT-III and α-ManII in CHO cells (GlycoMAb Technology)Enhanced survival of SCID-beige mice with A549-B34 NSCLC xenograftsMirschberger et al. 134 Humanized IgG1 Enhanced tumor inhibition in A549 orthotopic mouse models over WT-huMab-HER3 Palivizumab-NRespiratory syncytial virus (RSV)afucosylatedTransgenic N. benthamiana with RNAi to eliminate the expression of plant specific xylosyl and fucosyl transferase genesEnhanced RSV protection in rat models over fucosylated counterpart(d)…”

Section: Enhanced Adcc Activities By Afucosylated Antibodies In In VImentioning

confidence: 99%

The “less-is-more” in therapeutic antibodies: Afucosylated anti-cancer antibodies with enhanced antibody-dependent cellular cytotoxicity

Pereira

Chan

Lin

et al. 2018

mAbs

255

211

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Therapeutic monoclonal antibodies are the fastest growing class of biological therapeutics for the treatment of various cancers and inflammatory disorders. In cancer immunotherapy, some IgG1 antibodies rely on the Fc-mediated immune effector function, antibody-dependent cellular cytotoxicity (ADCC), as the major mode of action to deplete tumor cells. It is well-known that this effector function is modulated by the N-linked glycosylation in the Fc region of the antibody. In particular, absence of core fucose on the Fc N-glycan has been shown to increase IgG1 Fc binding affinity to the FcγRIIIa present on immune effector cells such as natural killer cells and lead to enhanced ADCC activity. As such, various strategies have focused on producing afucosylated antibodies to improve therapeutic efficacy. This review discusses the relevance of antibody core fucosylation to ADCC, different strategies to produce afucosylated antibodies, and an update of afucosylated antibody drugs currently undergoing clinical trials as well as those that have been approved.

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SuperiorIn vivoEfficacy of Afucosylated Trastuzumab in the Treatment of HER2-Amplified Breast Cancer

Cited by 199 publications

References 37 publications

Rapid screening of IgG quality attributes – effects on Fc receptor binding

Rapid screening of IgG quality attributes – effects on Fc receptor binding

Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

The “less-is-more” in therapeutic antibodies: Afucosylated anti-cancer antibodies with enhanced antibody-dependent cellular cytotoxicity

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